Protein Solubility
==================

Procedure
---------

1. Pellet 3 mL of culture in a 1.5 mL eppendorf tube.
2. Rinse pellet twice with 1 mL of 10 mM Tris, pH 7.5, 0.15 M NaCl.
3. Resuspend in 150 uL of 10 mM Tris, pH 7.5, 0.15 M NaCl.
4. Subject to 10 pulses of sonication, with a minimum power setting and 80% duty cycle.
5. Centrifuge sonicant at 14,000 G for 15 minutes.
6. Remove supernatant.
7. Wash pellet twice with 1 mL of 10 mM Tris, pH 7.5, 0.15 M NaCl.
8. Resuspend in final volume of 150 μL.
9. Remove a 5 uL aliquot of the pellet and supernatant.
10. Mix with 5 μL of SDS buffer containing DTT.
11. Heat for 15 minutes at 100 degrees in a thermocycler.
12. Resolve protein using a 12.5% acrylamide gel, stain with Coomassie.
13. Fix image gel using densiometry, allowing analysis of soluble and insoluble fraction.

Notes and Cautions
------------------

- Can use internal standard of BSA at 2 mg/mL.